Team:Western Canada/Parts
Part Table
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| Part Name | Part Number | Description | Source | Design Considerations |
| SpyCatcher-CsgA | BBa_K3316004 | Coding sequence of fusion protein product containg the SpyCatcher moiety linked by a 13 amino acid GS linker to the major curlin fibre precursor subunit, CsgA. Sequence is proceeded by a 6-His tag. | The CsgA sequence was derived from the Escherichia coli K12 genome. The SpyCatcher moiety is endogenous to Streptococcus pyogenes. | To help purify the fusion protein, we added a 6-His tag at the N-terminus of the protein. The choice of linker is to minimize constraints in conformation in either portion of the fusion protein product due to sterics. |
| CsgA-SpyCatcher | BBa_K3316005 | Coding sequence of fusion protein product containg the SpyCatcher moiety linked by a 13 amino acid GS linker to the major curlin fibre precursor subunit, CsgA. Sequence is proceeded by a 6-His tag. | The CsgA sequence was derived from the Escherichia coli K12 genome. The SpyCatcher moiety is endogenous to Streptococcus pyogenes. | To help purify the fusion protein, we added a 6-His tag at the N-terminus of the protein. The choice of linker is to minimize constraints in conformation in either portion of the fusion protein product due to sterics. |
| SpyTag-Cutinase | BBa_K3316006 | Fusion protein product containg the SpyTag moiety linked by a 12 amino acid GS linker to Cutinase (CueO), a cuticula degrading hydrolase from the fungus Fusarium solani pisi. The coding sequence is preceded by the 5' UTR and first 47 amino acids of the E. coli type III secretion system protein flagellin, FliC47 to promote extracellular transport of the protein product. A 6-His tag was added at the C-terminus of Cutinase to facilitate protein isolation. | The FliC 5'UTR and peptide sequence sequence was derived from the Escherichia coli K12 genome. The SpyTag moiety is endogenous to Streptococcus pyogenes. The cutinase enzyme is endogenous to Fusarium solani pisi. | To help purify the fusion protein, we added a 6-His tag at the N-terminus of the protein. The choice of linker is to minimize constraints in conformation in either portion of the fusion protein product due to sterics. |
| Cutinase-SpyTag | BBa_K3316007 | Fusion protein product containg the SpyTag moiety linked by a 12 amino acid GS linker to Cutinase (CueO), a cuticula degrading hydrolase from the fungus Fusarium solani pisi. The coding sequence is preceded by the 5' UTR and first 47 amino acids of the E. coli type III secretion system protein flagellin, FliC47 to promote extracellular transport of the protein product. A 6-His tag was added at the C-terminus of Cutinase to facilitate protein isolation. | The FliC 5'UTR and peptide sequence sequence was derived from the Escherichia coli K12 genome. The SpyTag moiety is endogenous to Streptococcus pyogenes. The cutinase enzyme is endogenous to Fusarium solani pisi. | To help purify the fusion protein, we added a 6-His tag at the N-terminus of the protein. The choice of linker is to minimize constraints in conformation in either portion of the fusion protein product due to sterics. |
| SpyTag-Laccase | BBa_K3316008 | Fusion protein product containg the SpyTag moiety linked by a 12 amino acid GS linker to E. coli K12 Lacccase (CueO), a copper-centric enzyme that catalyzes the oxidation of polyphenolic compounds. The coding sequence is preceded by the 5' UTR and first 47 amino acids of the E. coli type III secretion system protein flagellin, FliC47 to promote extracellular transport of the protein product. A 6-His tag was added at the C-terminus of Cutinase to facilitate protein isolation. | The FliC 5'UTR and peptide sequence was derived from the Escherichia coli K12 genome. The SpyTag moiety is endogenous to Streptococcus pyogenes. The laccase enzyme is endogenous to Escherichia coli K12. | To help purify the fusion protein, we added a 6-His tag at the N-terminus of the protein. The choice of linker is to minimize constraints in conformation in either portion of the fusion protein product due to sterics. |
| Laccase-SpyTag | BBa_K3316009 | Fusion protein product containing E. coli K12 Lacccase (CueO), a copper-centric enzyme that catalyzes the oxidation of polyphenolic compounds, linked by a 12 amino acid GS linker to the SpyTag moiety. The coding sequence is preceded by the 5' UTR and first 47 amino acids of the E. coli type III secretion system protein flagellin, FliC47 to promote extracellular transport of the protein product. A 6-His tag was added at the C-terminus of Cutinase to facilitate protein isolation. | The FliC 5'UTR and peptide sequence was derived from the Escherichia coli K12 genome. The SpyTag moiety is endogenous to Streptococcus pyogenes. The laccase enzyme is endogenous to Escherichia coli K12. | To help purify the fusion protein, we added a 6-His tag at the N-terminus of the protein. The choice of linker is to minimize constraints in conformation in either portion of the fusion protein product due to sterics. |
| FliC 5’UTR + 47aa | BBa_K3316003 | Escherichia coli flagellin secretion system signal sequence | Sequence derived from the E. coli type III flagellar secretion system, which is used by bacteria for export of unfolded FliC flagellin monomers. Green et al. (2019) propose a method, in which an E. coli strain with several gene deletions can export enzymes tagged with an N-terminal sequence of FliC19. The sequence contains the 5'UTR region and the first 47 amino acids of the FliC47 gene from E. coli K12. | n/a |
| Major Curlin Subunit Precursor - E coli K12(CsgA) | BBa_K3316001 | Major Curlin Subunit Precursor from Escherichia coli K12(CsgA) | Sequence derived Escherichia coli K12 | n/a |
| GS linker | BBa_K3316002 | Flexible 13 amino acid glycine-serine linker. | n/a | n/a |